人类免疫缺陷病毒口腔黏膜渗出液质控样本的制备  

Preparation of quality control samples for human immunodeficiency virus in oral mucosal transudate

作　　者：马仲慧 宫赛赛 裴丽健[2] 赫晓霞[2] 程焕义 蒋岩[2] 肖瑶[2] 潘品良[2] 邢文革[2] MA Zhong-hui;GONG Sai-sai;PEI Li-jian;HE Xiao-xia;CHENG Huan-yi;JIANG Yan;XIAO Yao;PAN Pin-liang;XING Wen-ge(Department of Infectious and Endemic Disease Control,Beijing Fangshan District Center for Disease Control and Prevention,Beijing 102488,China;不详)

机构地区：[1]北京市房山区疾病预防控制中心传染病与地方病控制科,北京102488 [2]中国疾病预防控制中心,性病艾滋病预防控制中心,参比实验室,北京102206

出　　处：《中国生物制品学杂志》2020年第7期788-793,共6页Chinese Journal of Biologicals

基　　金：中美艾滋病防治合作项目。

摘　　要：目的探讨人类免疫缺陷病毒(human immunodeficiency virus,HIV)口腔黏膜渗出液(oral mucosal transudate,OMT)质控样本的制备方法,以用于制备HIV OMT快速检测(rapid diagnostic test,RDT)试剂方法学评价的评价盘。方法用6家HIV OMT RDT试剂(编号A^F)样本稀释液(编号D1~D6)分别对1份抗-HIV阳性血浆样本进行倍比稀释(21~220),采用A试剂检测,测定相应的GOD(gold optical density)值,确定各稀释液的线性范围和最适稀释度;根据最适稀释度,用6种稀释液分别对抗-HIV阳性血浆样本进行稀释并分装,评价稀释液的均一性和稳定性,选出最适稀释液;用最适稀释液倍比稀释(21~220)4个系列混合阳性血浆样本,分别用A^F试剂检测,确定最适稀释度;用最适稀释液分别稀释40份抗-HIV阳性和40份HIV阴性血浆样本至最适稀释度制备质控样本,A试剂检测。结果稀释液D1~D6的线性范围分别为9~14、8~13、8~15、8~13、7~14、7~12(P<0.05),最适稀释度为210。均一性评价中,D4稀释后样本的CV<20%;稳定性评价中,37℃放置1~15 d,D1、D3稀释后样本的CV<20%,室温放置1~15 d,D1、D4、D5稀释后样本的CV<20%,2~8℃放置1~30 d,D1、D3、D4稀释后样本的CV<20%,-20℃放置1~30 d,D6稀释后样本的CV<20%;选择D1为最适稀释液。随着倍比稀释次数增加,A试剂、B、C、E、F的GOD值或显色强度先增加后降低,试剂D的GOD值逐渐降低。4种稀释系列满足A^F试剂线性范围的稀释度分别为29、210;28、29、210;210;28、29、210,从中选出210为最适稀释度。利用最适稀释液和稀释度制备的多份阳性质控样本的平均GOD值为5.82,标准差为3.10;阴性质控样本GOD值均为0。结论确定用于制备HIV OMT质控样本的最适稀释液为D1,最适稀释度为210。制备方法合适,可用于制备HIV OMT RDT评价盘。Objective To investigate the preparation method of quality control samples for human immunodeficiency virus(HIV)in oral mucosal transudate(OMT)and prepare the panels for methodological evaluation on the kit of rapid diagnostic test(RDT)for HIV OMT.Methods Six sets of serially diluted samples were prepared by doubly diluting an anti-HIV positive plasma sample from a dilution of 21 to 220 with sample diluents(marked as D1 to D6)from six HIV OMT RDT kits(marked as reagents A to F)separately and measured for gold optical density(GOD)with reagent A to determined the linear ranges of various diluents and the optimal dilution.The anti-HIV positive plasma sample was diluted with the six diluents to the optimal dilution and filled to evaluate the homogeneity and stability of diluents,based on which the optimal diluent was screened.Four series of mixed positive plasma samples were doubly diluted with the optimal diluent to the dilutions of 21~220,and determined with kits A^F separately to optimize the dilution.Forty anti-HIV positive and 40 HIV-negative plasma samples were diluted with the optimal diluent to the optimal dilution,and the prepared quality control samples were determined by kit A.Results The linear ranges of diluents D1~D6 were 9~14,8~13,8~15,8~13,7~14 and 7~12 respectively,while the optimal dilution was 210.In the homogeneity test,the CV of test results of sample diluted with D4 was less than 20%.However,in the stability test,the CVs of test results of samples diluted with D1 and D3 after storage at 37℃for 1~15 d,with D1,D4 and D5 after storage at room temperature for 1~15 d,with D1,D3 and D4 after storage at 2~8℃for 1~30 d and with D6 after storage at-20℃for 1~30 d were less than 20%.D1 was selected as the optimal diluent.With the increasing dilution,the GODs or chromogenic strengths of kits A,B,C,E and F increased at first then decreased,while the GOD of kit D decreased gradually.The dilutions of samples in series 1 within the linear range of kits A^F were 29 and 210,while in series 2 were 28,29 and

关 键 词：人类免疫缺陷病毒 口腔黏膜渗出液 质控样本 快速检测 稀释液 稀释度 

分 类 号：R373.9[医药卫生—病原生物学] R392-33[医药卫生—基础医学]